Molecular characterization of ocular dirofilariasis: A case report of Dirofilaria immitis in south-eastern Iran

Background: Dirofilariasis is a zoonotic parasitic infection transmitted from animals to humans by culicid mosquitoes. Although the disease can be caused by Dirofilaria spp. including Dirofilaria immitis and Dirofilaria repens, human ocular dirofilariasis due to D. immitis is relatively rare in the world. This study was aimed to present a case of ocular dirofilariasis caused by D. immitis in southeastern Iran. Case presentation: A nematode extracted from the right eye of a 69-year-old man referred with clinical symptoms including itching and redness was examined. After the morphometric analysis, Dirofilaria parasite was detected. Afterwards, a piece of worm body was cut and DNA was extracted and a 680-bp gene fragment amplification and nucleotide sequencing were performed. Phylogenetic analysis revealed a D. immitis roundworm as the causative agent of infection. The patient was treated with antibiotics and corticosteroid and followed up for 1 month. Conclusion: The present study provides the second report on ocular dirofilariasis caused by D. immitis isolated from a human in southeast Iran. Based on the available evidence, dirofilariasis in dogs has significantly increased in endemic areas such as Iran. Therefore, physicians should be aware of such zoonotic nematodes so as to take proper and timely action and treatment against the disease. © 2020 The Author(s)

The immunity of the recombinant prokaryotic and eukaryotic subunit vaccines against cutaneous leishmaniasis

Leishmaniasis counts as one of the most neglected tropical diseases. Despite the amount of research perceived in this field, no fully effective and approved vaccine against this disease is yet available in humans. In this study, LACK and KMP11 antigens were constructed simultaneously by recombinant methods in prokaryotic and eukaryotic expression systems and were compared and assessed along with the CpG adjuvant in BALB/c mice. In the prokaryotic method, LACK and KMP11 protein gene sequences were synthesized in pET28a-TEV vector. In order to extract these two proteins after expression, His-tag and S-tag sequences were added to the constructs, respectively for LACK and KMP11. The pET28a-TEV-LACK/KMP11 construct was transformed into Escherichia coli, and the inserts were verified by Colony PCR. Pure proteins were verified by western blot, and groups of BALB/c mice were injected with the created prokaryotic recombinant proteins along with an ODN CpG adjuvant. In the eukaryotic method, antigen sequences were constructed in the pLEXSY-neo 2.1 vector, E.coli Top10 strain was cloned in the bacteria, and after being linearized were transfected into Leishmania tarentolae genome. After recombinant strains were selected, they were verified by molecular methods. After the extraction and purification of the protein using the method above, groups of mice were injected with the recombinant antigens and ODN CpG adjuvant. Eukaryotic subunit vaccines showed more effective immunization compared with prokaryotic vaccines and caused an immune system shift towards Th1 and protection. Protein expression in L. tarentolae by the constructs created in this host contains Post-Translational Modifications. The constructed protein will be significantly similar to eukaryotic proteins, considering that they are identical epitopes. More comprehensive studies aiming to improve the effectiveness of this vaccine are being conducted to improve immune profiles and immunological memory stimulation in future designs. © 2021 Elsevier Lt

High Frequency of Diarrheagenic Escherichia coli in HIV-Infected Patients and Patients with Thalassemia in Kerman, Iran

This study was conducted on patients with thalassemia and HIV-infected patients to determine the frequency of diarrheagenic Escherichia coli in Kerman, Iran. We analyzed 68 and 49 E coli isolates isolated from healthy fecal samples of patients with thalassemia and HIV-infected patients, respectively. The E coli isolates were studied using a multiplex polymerase chain reaction to identify the enterotoxigenic E coli (ETEC), enterohemorrhagic E coli (EHEC), and enteropathogenic E coli (EPEC) groups. Statistical analysis was carried out to determine the correlation of diarrheagenic E coli between HIV-infected patients and patients with thalassemia using Stata 11.2 software. The frequency of having at least 1 diarrheagenic E coli was more common in patients with thalassemia (67.64%) than in HIV-infected patients (57.14%; P = .25), including ETEC (67.64% versus 57.14%), EHEC (33.82% versus 26.53%), and EPEC (19.11% versus 16.32%). The results of this study indicate that ETEC, EHEC, and EPEC pathotypes are widespread among diarrheagenic E coli isolates in patients with thalassemia and HIV-infected patient

Prevalence of Trypanosoma evansi in camels using molecular and parasitological methods in the southeast of Iran, 2011

Surra is caused by infection with the protozoal parasite, Trypanosoma evansi. This parasite was transmitted mechanically by biting flies which is widespread in camels in the world. The aim of this study is to determine the prevalence of T. evansi in camels in Rafsanjan, Kerman province, southeast of Iran. In this study, 95 suspected camels were randomly selected in 2011. Blood samples were taken from deep blood vessels. Thin and thick blood smears were prepared in laboratory. Blood smears were stained by Giemsa and studied under a light microscope. The positive blood samples were also used for further molecular analysis. Data were analyzed using SPSS 17.0 software and P ≤ 0.05 was considered as statistical difference. A total of 95 camels were examined for infection with T.evansi using parasitological and molecular methods. The overall prevalence of infection was 2.1 %. It was found that the frequency of infection was significantly higher (P 6 years old than the corresponding younger camels. However, there was no significant difference when the gender was considered. PCR technique confirmed the two infected cases were T. evansi. Results of the present study indicated that surra is present in Rafsanjan county, Kerman province in an infection rate of 2.1 % in camels. To our knowledge, this is the first study reported from this province. Further investigations are needed to focus on vectors and to evaluate the risk factors

Epidemiological and molecular studies on Echinococcus granulosus from free-roaming dogs in Southeast Iran

Background and Aim: Cystic echinococcosis (CE), as a major zoonotic helminth infection, imposes remarkable socioeconomic burden on many endemic countries across the world, including Iran. Due to the high importance of free-roaming dogs in the transmission of CE, epidemiological and molecular studies in this type of hosts are required in the endemic regions. This study aimed to investigate the epidemiology and genotyping of Echinococcus granulosus isolated from stray dogs in Kerman, Southeast Iran. Materials and Methods: Eighty-four samples were isolated from stray dogs in the city and suburbs of Kerman in coordination with the health authorities and the municipality office for rabies control and dog population management. Dog demographic data, including age and sex were collected. The worm was isolated by necropsy and genomic DNA was extracted and partial cytochrome c oxidase subunit 1 gene was amplified using specific primers. Phylogenetic and Templeton-Crandall-Sing (TCS) network analyses were carried out on the sequence data. Results: The overall prevalence of CE in the surveyed dogs was 10.7 (9/84 cases). Out of 84 stray dogs, 33 (39.3) and 51 (60.7) cases were male and female, respectively. There was not a statistically significant difference between the infection and gender of dogs. However, infection is shown more in dogs under one year of age with a statistically significant difference (p<0.05). The results of molecular studies indicated E. granulosus G1 genotype for all isolates. The high presence of free-roaming dogs in urban and peri-urban areas and high frequency of parasite in this animal is a risk factor for humans in the region. Haplotype sequence analysis on the dog isolates revealed a close relationship with other E. granulosus isolates in Kerman. Conclusion: The findings of this study provide evidence-based data about the epidemiological and molecular characteristics of CE in dog definitive hosts of Southeast Iran. Further studies are required to understand the prevalence and parasite genotypes in dogs in Iran. © 202

New foci of zoonotic cutaneous leishmaniosis due to Leishmania major in the northeastern Iran cities of Sabzevar and Neghaab

Cutaneous leishmaniosis (CL) is a major public health challenge in Iran. Overall, 18 provinces out of 31 of the country's provinces are in the endemic areas. The objective of the present study was to determine the presence of CL species in Sabzevar and Neghaab cities, Khorasan Razavi province, northeastern Iran. Overall, 280 samples were taken from suspected individuals referred to Sabzevar's and Neghaab's health centers during 2014-2016. Smear preparation, case detection and further identification by partial amplification of 7SL RNA and high-resolution melting curve analysis were done for all samples, followed by randomly polymerase chain reaction (PCR)-sequencing confirmation. Based on findings, the most infection rate was found in males with the age groups of 20-30 in both districts. All samples were characterized as Leishmania major, except one isolate in Neghaab closely related to Leishmania tropica. Various risk factors play roles in creating new foci of zoonotic CL (ZCL) caused by L. major in Sabzevar and Neghaab in northeastern Iran. Reports of the prevalence of CL in new foci make serious concerns about the incidence of cases and expansion of disease to the neighboring areas. Further measures are essential to control the disease in the regions

Dirofilariosis caused by Dirofilaria immitis in the south of Kerman province, Iran

Background: Dirofilariosis due to Dirofilaria immitis is endemic in various areas of Iran. Domestic dogs are the main reservoirs and represent a major potential infection source for the vector and humans. Objective: This study aimed to explore the prevalence of dirofilariosis due to D.immitis and its public health importance in domestic dogs in the Jiroft district, south of Kerman province, Iran, by serological and parasitological methods. Methods: This descriptive study was carried out as a cross-sectional investigation. A questionnaire was completed for 100 domestic dogs from May 2017 to February 2018 and recorded their age, sex, and clinical features. Also, we used enzyme-linked immunosorbent assay (ELISA) to identify antigens of heartworms in the bloodstream, with 98 sensitivity and 100 specificity, and parasitological techniques (Knott's test) to detect microfilariae in canine blood in Jiroft district, south of Kerman province, Iran. Results: Overall, 10 (10) and 4 (4) domestic dogs were infected as confirmed by ELISA and modified Knott's tests, respectively. The rate of occult infections in the ELISA test than Knott's test was 60. No significant difference was found between dirofilariosis and gender. In contrast, there was a significant difference between dirofilariosis infection and age (P < 0.05). Conclusion: The present findings could help understand the epidemiological aspects of D. immitis for future control programs and take appropriate preventive and therapeutic strategies against the disease. © 2021 Elsevier Lt

Leishmania tropica in Stray Dogs in Southeast Iran

BACKGROUND: Cutaneous leishmaniasis (CL) caused by Leishmania tropica is endemic in Kerman, southeastern Iran. While dogs have long been implicated as the main domestic reservoirs of L. infantum, etiological agent of zoonotic visceral leishmaniasis (ZVL), they can also carry L. tropicainfection. The objective of the present study was to determine molecular identity and to evaluate histopathological changes due to CL in dogs in a well-known focus of anthroponotic CL (ACL) in Kerman, southeastern Iran. METHODS: This study was carried out in three prospective series from 1994 to 2013 on dogs. Tissue samples were taken from 471 stray dogs. Pathological specimens including skin, spleen, liver and lymph nodes were prepared for paraffin blocks, sectioning and staining for further histopathological examination. PCR amplification of kDNA was performed to identify the causative agent and sequencing. Overall, two out of 471 stray dogs were infected with L. tropica. Hyperplasia of red pulp by the proliferation of histiocytes, lymphocytes, plasma cells and cytoplasm of histiocytes collection of amastigotes was noted. RESULTS: Based on the results of PCR products and sequencing analysis, the parasites isolated from the lesions of two dogs were characterized as L. tropica, corresponding to a band of 830 bp. CONCLUSION: This finding revealed infection with L. tropica in stray dogs in the city and suburbs of Kerman. This information is essential for public health concerns and planning effective future control programs. The role of dogs as potentional reservoir in the epidemiology of ACL needs further investigation

Linguatula serrata in cattle in southeastern Iran: Epidemiological, histopathological and phylogenetic profile and its zoonotic importance

This study aimed to determine the prevalence, histopathological observations, and phylogenetic analysis of L. serrata in cattle and its potential zoonotic and public health implications in southeastern Iran.The cross-sectional study was performed in cattle in southeastern Iran. Lymph nodes were collected from each cattle and examined by parasitological and histopathological techniques. A binary logistic regression and chi-square tests were implemented to analyze the data. Genomic DNA was randomly extracted from the nymphal stages of Linguatula isolates. Further characterization and phylogenetic relationships were done using two primers for amplification of partial DNA fragments of 18 s rRNA and cytochrome C oxidase subunit 1 (cox1), respectively.The results showed that 64 cattle of the total 404 were infected with L. serrata. There was no significant difference between linguatulosis infection and gender, while age was significantly different (P < 0.05). The cyst-like spaces containing the longitudinal and transverse sections of the L. serrata nymphs were surrounded by granulomatous reactions. The higher nucleotide variation in the cox1 region was supported by estimating the evolutionary divergence between L. serrata isolates and other Linguatula records of ruminants in Iran. The phylogenetic tree confirmed the close evolutionary relationships among all reported records of L. serrata in Iran.The high prevalence of linguatulosis caused by L. serrata declares the existence of a potential risk of FBPs for humans in southeastern Iran. This condition can advance more serious public health problems and requires a comprehensive control program and treatment strategies to prevent the disease. © 2020 Elsevier B.V